- Molecular identification of carnosine synthase as ATP-grasp domain-containing protein 1 (ATPGD1)
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Carnosine (β-alanyl-L-histidine) and homocarnosine (γ-aminobutyryl-L-histidine) are abundant dipeptides in skeletal muscle and brain of most vertebrates and some invertebrates. The formation of both compounds is catalyzed by carnosine synthase, which is thought to convert ATP to AMP and inorganic pyrophosphate, and whose molecular identity is unknown. In the present work, we have purified carnosine synthase from chicken pectoral muscle about 1500-fold until only two major polypeptides of 100 and 90 kDa were present in the preparation. Mass spectrometry analysis of these polypeptides did not yield any meaningful candidate. Carnosine formation catalyzed by the purified enzyme was accompanied by a stoichiometric formation, not of AMP, but of ADP, suggesting that carnosine synthase belongs to the "ATP-grasp family" of ligases. A data base mining approach identified ATPGD1 as a likely candidate. As this protein was absent from chicken protein data bases, we reconstituted its sequence from a PCR-amplified cDNA and found it to fit with the 100-kDa polypeptide of the chicken carnosine synthase preparation. Mouse and human ATPGD1 were expressed in HEK293T cells, purified to homogeneity, and shown to catalyze the formation of carnosine, as confirmed by mass spectrometry, and of homocarnosine. Specificity studies carried out on all three enzymes were in agreement with published data. In particular, they acted with 15-25-fold higher catalytic efficiencies on β-alanine than on γ-aminobutyrate. The identification of the gene encoding carnosine synthase will help for a better understanding of the biological functions of carnosine and related dipeptides, which still remain largely unknown.
- Drozak, Jakub,Veiga-da-Cunha, Maria,Vertommen, Didier,Stroobant, Vincent,Van Schaftingen, Emile
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- Solid-phase peptide synthesis of dipeptide (histidine-β-Alanine) as a chelating agent by using trityl chloride resin, for removal of Al3+, Cu2+, Hg2+ and Pb2+: Experimental and theoretical study
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Solid-phase peptide synthesis of dipeptide (histidine-β-Alanine) as a chelating agent examined by common N-9-fluorenylmethyloxycarbonyl-N-Trityl-L-histidine and tert-butyloxycarbonyl-β-Alanine-OH amino acid derivatives. Trityl chloride resin was used as a carrier resin. The molecular structure of the dipeptide was definite by using different methods such as ultraviolet visible (UV-Vis), Fourier transform infrared (FTIR), proton (1H) nuclear magnetic ressonance (NMR) and liquid chromatography-mass spectrometry (LC-MS) and the chelating property of synthesized dipeptide was investigated for removing of metal ions Al3+, Cu2+, Hg2+ and Pb2+ in vitro. In addition, the pharmacological and biological activities of dipeptide were examined by prediction of activity spectra for substances (PASS) program.
- Rahimi, Rahmatollah,Khosravi, Maryam,Tehrani, Mohammd H. H.,Rabbani, Mahboubeh,Safavi, Ebrahim
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Read Online
- Carnosine intermediate and preparation and application of carnosine thereof
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The invention provides a carnosine intermediate and preparation and application of carnosine thereof, and belongs to the technical field of medical intermediates. The preparation method of the carnosine intermediate comprises the following steps: carrying out acylation reaction on L-histidine and tert-butyl cyanoacetate under the action of a catalyst to prepare the carnosine intermediate; wherein isonicotinic acid and 4-chloropyrimidine are added as catalytic assistants. When the carnosine intermediate is prepared, isonicotinic acid and 4-chloropyrimidine are used as auxiliaries, and the yield of the obtained carnosine intermediate is greatly improved. The method for preparing carnosine from the carnosine intermediate prepared by the method comprises the step of adding a catalyst into the carnosine intermediate for catalytic hydrogenation reduction to obtain the carnosine. The composition prepared from the obtained carnosine, theanine and loganic acid has the effects of resisting oxidation and whitening.
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Paragraph 0039; 0040
(2022/03/27)
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- METHODS AND COMPOSITIONS FOR RAPIDLY DECREASING EPIGENETIC AGE AND RESTORATION OF MORE YOUTHFUL FUNCTION
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Disclosed are methods and compositions of reducing the epigenetic age of mammalian organism, especially an adult human of geriatric age. The methods provide for the proliferation of endogenous stem cells using mitochondrial fusion and a UCP2 blocker or other stimulants; supplying stem cells with nutrition to prevent cell cycle arrest; and removal of senescent somatic cells using senolytic treatments. The proliferation of endogenous neural stem cells after plaque removal for the treatment of Alzheimer's is also disclosed.
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- A green-by-design bioprocess for l-carnosine production integrating enzymatic synthesis with membrane separation
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l-Carnosine (l-Car, β-alanyl-l-histidine) is a bioactive dipeptide with important physiological functions. Direct coupling of unprotected β-Ala (β-alanine) with l-His (l-histidine) mediated by an enzyme is a promising method for l-Car synthesis. In this study, a new recombinant dipeptidase (SmPepD) from Serratia marcescens with a high synthetic activity toward l-Car was identified by a genome mining approach and successfully expressed in Escherichia coli. Divalent metal ions strongly promoted the synthetic activity of SmPepD, with up to 21.7-fold increase of activity in the presence of 0.1 mM MnCl2. Higher temperature, lower pH and increasing substrate loadings facilitated the l-Car synthesis. Pilot biocatalytic syntheses of l-Car were performed comparatively in batch and continuous modes. In the continuous process, an ultra-filtration membrane reactor with a working volume of 5 L was employed for catalyst retention. The dipeptidase, SmPepD, showed excellent operational stability without a significant decrease in space-time yield after 4 days. The specific yield of l-Car achieved was 105 gCar gcatalyst-1 by the continuous process and 30.1 gCar gcatalyst-1 by the batch process. A nanofiltration membrane was used to isolate the desired product l-Car from the reaction mixture by selectively removing the excess substrates, β-Ala and l-His. As a result, the final l-Car content was effectively enriched from 2.3% to above 95%, which gave l-Car in 99% purity after ethanol precipitation with a total yield of 60.2%. The recovered substrate mixture of β-Ala and l-His can be easily reused, which will enable the economically attractive and environmentally benign production of the dipeptide l-Car.
- Yin, Dong-Ya,Pan, Jiang,Zhu, Jie,Liu, You-Yan,Xu, Jian-He
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p. 5971 - 5978
(2019/11/14)
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- METHOD FOR PREPARING PEPTIDES
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The invention relates to a method for preparing peptides comprising the step of forming a peptide bond wherein at least one amino acid or peptide comprises a protecting group having a water-solubility enhancing group, and said forming of a peptide bond is achieved while an amino acid or a peptide is bound to a solid phase. The invention further relates to peptides comprising a protecting group having a water-solubility enhancing group being bound to the amino group and an activated or free carboxyl group.
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Page/Page column 73
(2019/06/11)
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- METHOD FOR PRODUCING L-CARNOSINE DERIVATIVE AND L-CARNOSINE
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PROBLEM TO BE SOLVED: To provide a convenient method for producing a high-purity N-protected L-carnosine derivative and L-carnosine. SOLUTION: A production method includes reacting an acid halide (1) with an L-histidine derivative (2). (R1 and R2 are H or a protection group of an amino group; X is a halogen atom). (A TMS group is a trimethylsilyl group). SELECTED DRAWING: None COPYRIGHT: (C)2020,JPOandINPIT
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- METHOD FOR PREPARING PEPTIDES
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The invention relates to a method for preparing peptides comprising the step of forming a peptide bond wherein the carboxyl group of a first amino acid or first peptide is activated and an amino group of the first activated amino acid or first peptide is protected by a protecting group having a water-solubility enhancing group and the activated carboxyl group of the first amino acid or first peptide is reacted with an amino group of a second amino acid or second peptide wherein said carboxyl group of the first amino acid or first peptide is activated in the absence of the second amino acid or second peptide. The invention further relates to peptides comprising a protecting group having a water-solubility enhancing group being bound to the amino group and an activated or free carboxyl group.
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Page/Page column 66-67
(2019/06/11)
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- Synthetic method of L-carnosine
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The invention discloses a synthetic method of L-carnosine, and belongs to the technical field of organic matter synthesis. The method comprises the following steps: dissolving 3-chloropropionic acid in an organic solvent, and converting the 3-chloropropionic acid by a chloroformylation reagent to form corresponding 3-chloropropionyl chloride; condensing trimethylsilane protected L-histidine and the 3-chloropropionyl chloride to obtain a corresponding amide product; removing protection agents by using water or an alkaline solution to obtain an intermediate; carrying out aminolysis on the intermediate to obtain crude L-carnosine; and purifying the crude L-carnosine to obtain finished L-carnosine. The synthetic method has the advantages of low raw material consumption, short reaction steps, few wastes, high yield, obtaining of high-quality L-carnosine free from hydrazine, and meeting of industrial production demands.
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Paragraph 0005; 0032; 0033; 0034; 0035
(2017/08/28)
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- Concise Synthesis of Anserine: Efficient Solvent Tuning in Asymmetric Hydrogenation Reaction
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A concise synthesis of anserine and related compounds was accomplished by Et-DuPhos-Rh-catalyzed asymmetric hydrogenation of dehydrohistidine derivatives in 2,2,2-trifluoroethanol, which played a key role in improving the yield and selectivity.
- Yamashita, Megumi,Shimizu, Keita,Koizumi, Yasuaki,Wakimoto, Toshiyuki,Hamashima, Yoshitaka,Asakawa, Tomohiro,Inai, Makoto,Kan, Toshiyuki
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supporting information
p. 2734 - 2736
(2016/11/30)
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- Supramolecular interactions in sodium N-(ethoxythioxomethyl)-β- alaninate-water (4/6) crystal and its application in synthesis of L-carnosine
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Compound sodium N-(ethoxythioxomethyl)-β-alaninate (sodium 3-(ethoxycarbonothioyl)propanoate) was synthesized and characterized by IR, 1HNMR, ESI-HRMS and single crystalX-ray diffraction. Single crystalX-ray diffraction analysis showed that the title compound crystallized in the triclinic space group P-1 with cell parameters a = 10.142 (2) A, b = 13.738 (3) A, c = 15.751 (3) A, α= 72.937 (2)°, β = 78.694 (2)°, γ = 89.999 (2)°, V = 2053.4 (7) A3, Dc = 1.464 g cm-3, Z = 2. In the extended structure of sodium N-(ethoxythioxomethyl)-β-alaninate-water (4/6), (NaL)4· 6H2O [L = CH3CH2OC(S)NHCH2CH 2COO], ligands (Ls) are stabilized by intermolecular O-H···O, N-H···O, C- H···O and weak O-H···S and C-H···S linkages, which further consolidate the crystal packing, making the ligand chains stacked along [0-1 1], and intramolecular O-H···S hydrogen bond is also observed. Each Na atom is surrounded with six O atoms, forming an octahedron and mutually bonded as tetramers. These tetramers are linked through O atom bridges from water molecules, extending as layers in the ab plane. In addition, synthesis of β-alanyl dipeptides was developed with particular focus on the preparation of L-carnosine. Springer Science+Business Media New York 2012.
- Zhang, Shi-Jie,Xu, Feng,Yang, Wei-Ji,Hu, Wei-Xiao
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p. 1182 - 1189
(2013/02/22)
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- Kinetic analysis of L-carnosine formation by β-aminopeptidases
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The β,α-dipeptide L-carnosine occurs in high concentrations in long-lived innervated mammalian tissues and is widely sold as a food additive. On a large scale L-carnosine is produced by chemical synthesis procedures. We have established two aqueous enzymatic reaction systems for the preparation of L-carnosine using the dissolved bacterial β-aminopeptidases DmpA from Ochrobactrum anthropi and BapA from Sphingosinicella xenopeptidilytica as catalysts and investigated the kinetics of the enzymecatalyzed peptide couplings. DmpA catalyzed the formation of L-carnosine from C-terminally activated β-alanine derivatives (acyl donor) and L-histidine (acyl acceptor) in an aqueous reaction mixture at pH 10 with high catalytic rates (Vmax=19.2 mmol min-1 per mg of protein, k cat=12.9 s-1), whereas Vmax in the BapA-catalyzed coupling reaction remained below 1.4 mmol min-1 per mg of protein (k cat=0.87 s-1). Although the equilibrium of this reaction lies on the side of the hydrolysis products, the reaction is under kinetic control and L-carnosine temporarily accumulated to concentrations that correspond to yields of more than 50% with respect to the employed acyl donor. However, competing nucleophiles caused unwanted hydrolysis and coupling reactions that led to decreased product yield and to formation of various peptidic by-products. The substitution of l-histidine for L-histidine methyl ester as acyl acceptor shifted the pKa of the amino functionality from 9.25 to 6.97, which caused a drastic reduction in the amount of coupling by-products in an aqueous reaction system at pH 8.
- Heck, Tobias,Makam, Venkata S.,Lutz, Jochen,Blank, Lars M.,Schmid, Andreas,Seebach, Dieter,Kohler, Hans-Peter E.,Geuekea, Birgit
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experimental part
p. 407 - 415
(2010/06/11)
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- N-trifluoroacetyl-β-alanine in the synthesis of carnosine
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Conditions have been developed for the synthesis of N-trifluoroacetyl- β-alanine, N-tifluoroacetyl-β-alanyl chloride, and N-trifluoroacetyl-β-alanine 4-nitrophenyl ester. These compounds reacted with histidine methyl ester or sodium salt to give N-trifluoroacetyl-β- alanyl-l-histidine methyl ester CF3CONHCH2CH 2?CONHCH(CH2C3H3N 2)COOCH3 and N-trifluoroacetyl-β-alanyl-l-histidine CF3CONHCH2CH2CONHCH?(CH2C 3H3N2)COOH. Their hydrolysis with a solution of sodium hydroxide in aqueous ethanol, followed by acidification with trifluoroacetic acid, led to the formation of β-alanyl-l-histidine (l-carnosine).
- Cherevin,Zubreichuk,Popova,Gulevich,Knizhnikov
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p. 1576 - 1579
(2008/03/13)
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- Method for producing beta-alaninamides
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The invention relates to β-alaninamides of general formula (I), wherein: R1 represents hydrogen or C1-C6 alkyl which is optionally substituted by hydroxy, amino, carboxy, carbamoyl, methylmercapto, guanidino, or by optionally substituted aryl or heteroaryl; R2 represents hydrogen or R1 and R2, together, form a group of formula —(CH2)n, wherein n represents 3 or 4, and; R3 represents hydrogen, a negative charge compensated for by an equivalent of an inorganic or organic cation or represents C1-C6 alkyl. The β-alaninamides are produced without using an amino protective group by reacting the corresponding α-amino acid or the corresponding α-amino acid ester with a cyanoacetic acid to form a cyanoacetic acid amide and by a subsequent catalytic hydrogenation. The method is particularly suited for producing carnosine (βalanyl-L-histidine, R1=imidazol-4-yl-methyl, R2═R3═H), a naturally occurring dipeptide, which is used as a food additive having an antioxidant effect.
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- Process for preparing L-carnosine
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Process for preparing L-carnosine by reacting L-histidine and tetrahydro-1,3-thiazin-2,4-dione in an aqueous medium at a selected pH and temperature range followed by removal of carbon oxysulfide at acid pH, removal of water and crystallization of product wherein pH adjustments are made with substituted ammonium hydroxides and organic acids having pKa ≤3.75.
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